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红树林根际土壤中耐高盐苯酚降解菌的分离鉴定
曹宏明,龚斌,朱丽娟,潘贵妮,刘飞琪,李世盛
0
(广西北部湾海洋灾害研究重点实验室,广西 钦州 535011;北部湾大学海洋学院,广西 钦州 535011;广西北部湾海洋生物多样性养护重点实验室,广西 钦州 535011)
摘要:
苯酚对海洋生态环境具有较大的威胁,从红树林根际土壤中分离获得对环境具有高适应性的高效芳烃降解菌具有重要的意义。本研究对北部湾红树林根际土壤中的苯酚降解菌进行富集培养,经过分离纯化得到30株菌株,筛选得到5株菌株FGYS1、FGYS7、FGYS11、FGYB1和FGYB5,可耐受70 g/L NaCl的高盐环境,最高可降解1 500 mg/L的苯酚。采用形态观察、BOXPCR菌种多样性分析、16S rRNA序列分析等方法对菌株进行鉴定,均为假单胞菌属(Pseudomonas)。菌株FGYS1、FGYS7和FGYS11为同一种假单胞菌,与Pseudomonas extremaustralis 14-3TPseudomonas meridiana CMS 38T的16S rRNA序列相似性分别达到99.71%和99.64%。菌株FGYB1和FGYB5为同一种假单胞菌,分别与Pseudomonas veronii DSM 11331TPseudomonas extremaustralis 14-3T的16S rRNA序列相似性均达到99.64%。16S rRNA序列的系统发育树分析表明菌株FGYS1、FGYS7、FGYS11、FGYB1和FGYB5在进化上与P. veronii为同一个分支,而与假单胞菌属其他模式菌株的序列进化关系相距较远。菌株 FGYS1 降解苯酚的适宜温度为25~35 ℃、适宜pH为4~6、最高耐NaCl含量为60 g/L;菌株 FGYB1 降解苯酚的适宜温度为25~35 ℃、适宜pH为4~6、最高耐NaCl含量为100 g/L,两株菌株对pH、温度和盐度的适应范围较广。菌株FGYS1和FGYB1还可以同时降解邻苯二酚、对苯二酚和间苯二酚。本研究分离得到的苯酚降解菌在应对红树林芳烃污染和生态环境修复方面显示了较大的潜力。
关键词:  海洋生物学  红树林根际土壤  苯酚  微生物降解  环境因素
DOI:10.3969/J.ISSN.2095-4972.2021.02.001
基金项目:国家自然科学基金资助项目(31560727);广西自然科学基金资助项目(2018JJA130187,2020GXNSFDA238015);广西高校中青年教师基础能力提升项目资助项目(2017KY0778);广西北部湾海洋灾害研究重点实验室资助项目(2017ZD01);国家级大学生创新创业训练计划资助项目(201811607038);广西北部湾海洋生物多样性养护重点实验室自主课题资助项目(2021ZA01)
Isolation and identification of halotolerant and phenol-degrading bacterial strains from rhizosphere soil of mangrove
CAO Hong-ming,GONG Bin,ZHU Li-juan,PAN Gui-ni,LIU Fei-qi,LI Shi-sheng
(Guangxi Key Laboratory of Marine Disaster in the Beibu Gulf, Qinzhou 535011, China;School of Ocean Sciences, Beibu Gulf University, Qinzhou 535011, China;Guangxi Key Laboratory of Beibu Gulf Marine Biodiversity Conservation, Qinzhou 535011, China;School of Ocean Sciences, Beibu Gulf University, Qinzhou 535011, China)
Abstract:
Phenol is a great threat to marine ecological system, so it is of great significance to isolate phenol-degrading bacteria characterizing higher effectiveness and higher adaptation of marine environment. In present work, phenol-degrading bacteria were enriched and isolated from rhizosphere soil of mangrove around Beibu Gulf. A total of 30 strains were isolated and purified. As a result, five strains named FGYS1, FGYS7, FGYS11, FGYB1 and FGYB5 were obtained. It shows that they could tolerate 70 g/L high concentration of sodium chloride and degrade phenol up to 1 500 mg/L. They were identified by morphological observation, BOX-PCR diversity analysis,16S rRNA and gyrB gene sequence analysis. All of them were Pseudomonas. FGYS1, FGYS7 and FGYS11 are consistent in 16S rRNA gene sequence. The nucleotide sequence similarity of 16S rRNA gene to those of Pseudomonas extremaustralis 14-3T and Pseudomonas meridianaCMS 38T in the EzBioCloud data base was 99.71% and 99.64%, respectively. It is the same for the strains FGYB1 and FGYB5, the similarity of 16S rRNA gene to those of Pseudomonas veroniiDSM 11331T and Pseudomonas extremaustralis14-3T was both 99.64%, respectively. The phylogenetic tree analysis of 16S rRNA gene showed that FGYS1, FGYS7, FGYS11, FGYB1 and FGYB5 located in the same branch with P. veronii, but relatively far away from other model strains of Pseudomonas. The suitable growth conditions for phenol degradation by strain FGYS1 were 25-35 ℃, pH 4-6 and the sodium chloride concentration less than 60 g/L. While for strain FGYB1, the conditions for phenol degradation were 25-35 ℃, pH 4-6, and the sodium chloride concentration less than 100 g/L. FGYS1 and FGYB1 could also degrade catechol, hydroquinone and resorcinol simultaneously. These phenol-degrading bacteria exhibited potential utilization in the disposition of aromatic pollution and the restoration of environment in mangrove ecosystem.
Key words:  marine biology  rhizosphere soil of mangrove  phenol  microbial degradation  environmental factor

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